scholarly journals Yeast Actin-Related Protein ARP6 Negatively RegulatesAgrobacterium-Mediated Transformation of Yeast Cell

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Yumei Luo ◽  
Zikai Chen ◽  
Detu Zhu ◽  
Haitao Tu ◽  
Shen Quan Pan
Keyword(s):  
Genetic Engineering ◽  
Large Scale ◽  
Transformation Efficiency ◽  
Genetic Diseases ◽  
Negative Regulator ◽  
Yeast Cells ◽  
Scale Production ◽  
Related Protein ◽  
Cellular Mechanisms ◽  
Eukaryotic Organisms

The yeasts, includingSaccharomyces cerevisiaeandPichia pastoris, are single-cell eukaryotic organisms that can serve as models for human genetic diseases and hosts for large scale production of recombinant proteins in current biopharmaceutical industry. Thus, efficient genetic engineering tools for yeasts are of great research and economic values.Agrobacterium tumefaciens-mediated transformation (AMT) can transfer T-DNA into yeast cells as a method for genetic engineering. However, how the T-DNA is transferred into the yeast cells is not well established yet. Here our genetic screening of yeast knockout mutants identified a yeast actin-related proteinARP6as a negative regulator of AMT.ARP6is a critical member of the SWR1 chromatin remodeling complex (SWR-C); knocking out some other components of the complex also increased the transformation efficiency, suggesting thatARP6might regulate AMT via SWR-C. Moreover, knockout ofARP6led to disruption of microtubule integrity, higher uptake and degradation of virulence proteins, and increased DNA stability inside the cells, all of which resulted in enhanced transformation efficiency. Our findings have identified molecular and cellular mechanisms regulating AMT and a potential target for enhancing the transformation efficiency in yeast cells.

scholarly journals Cannabis sativa: From Therapeutic Uses to Micropropagation and Beyond

Plants ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2078
Author(s):  
Tristan K. Adams ◽  
Nqobile A. Masondo ◽  
Pholoso Malatsi ◽  
Nokwanda P. Makunga
Keyword(s):  
Tissue Culture ◽  
Genetic Engineering ◽  
Large Scale ◽  
Gene Editing ◽  
Cannabis Sativa ◽  
Scale Production ◽  
Indirect Organogenesis ◽  
Large Scale Production ◽  
Pharmaceutical Industries

The development of a protocol for the large-scale production of Cannabis and its variants with little to no somaclonal variation or disease for pharmaceutical and for other industrial use has been an emerging area of research. A limited number of protocols have been developed around the world, obtained through a detailed literature search using web-based database searches, e.g., Scopus, Web of Science (WoS) and Google Scholar. This article reviews the advances made in relation to Cannabis tissue culture and micropropagation, such as explant choice and decontamination of explants, direct and indirect organogenesis, rooting, acclimatisation and a few aspects of genetic engineering. Since Cannabis micropropagation systems are fairly new fields, combinations of plant growth regulator experiments are needed to gain insight into the development of direct and indirect organogenesis protocols that are able to undergo the acclimation stage and maintain healthy plants desirable to the Cannabis industry. A post-culture analysis of Cannabis phytochemistry after the acclimatisation stage is lacking in a majority of the reviewed studies, and for in vitro propagation protocols to be accepted by the pharmaceutical industries, phytochemical and possibly pharmacological research need to be undertaken in order to ascertain the integrity of the generated plant material. It is rather difficult to obtain industrially acceptable micropropagation regimes as recalcitrance to the regeneration of in vitro cultured plants remains a major concern and this impedes progress in the application of genetic modification technologies and gene editing tools to be used routinely for the improvement of Cannabis genotypes that are used in various industries globally. In the future, with more reliable plant tissue culture-based propagation that generates true-to-type plants that have known genetic and metabolomic integrity, the use of genetic engineering systems including “omics” technologies such as next-generation sequencing and fast-evolving gene editing tools could be implemented to speed up the identification of novel genes and mechanisms involved in the biosynthesis of Cannabis phytochemicals for large-scale production.

scholarly journals Development of plant regeneration and Agrobacterium tumefaciens-mediated transformation methodology for Physalis pruinosa

2018 ◽  
Author(s):  
Kerry Swartwood ◽  
Joyce Van Eck
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Plant Regeneration ◽  
Genetic Engineering ◽  
Large Scale ◽  
Gene Editing ◽  
Fluorescent Protein ◽  
Pcr Analysis ◽  
Marker Genes ◽  
Scale Production ◽  
Hypocotyl Explants

AbstractPhysalis pruinosa, also known as groundcherry, produces a small, yellow, highly nutritious edible fruit that is enveloped by a papery husk. In order for the potential of large-scale production of P. pruinosa fruit to be realized, undesirable characteristics, such as an unmanageable, sprawling growth habit and extensive fruit drop, need to be improved by exploiting approaches available through plant breeding, genetic engineering, and gene editing. In this study, we established plant regeneration and Agrobacterium tumefaciens-mediated methods to allow application of genetic engineering and gene editing of P. pruinosa. Cotyledon and hypocotyl explants from 7 – 8-day-old in vitro-grown seedlings were assessed for plant regeneration. Explants were cultured for 2 weeks on a Murashige and Skoog salts-based medium that contained 2 mg/L zeatin followed by transfer to medium containing 1 mg/L zeatin. Only hypocotyl explants regenerated shoots. Hypocotyl explants were infected with Agrobacterium tumefaciens strain AGL1 containing the pJL33 binary vector that has the green fluorescent protein (GFP) reporter and neomycin phosphotransferase II (nptII) selectable marker genes. After cocultivation, explants were cultured on selective plant regeneration medium that contained 50, 100, 200, 250, and 300 mg/L kanamycin to determine the most effective level for efficient recovery of transgenic lines. Based on rooting of regenerated shoots on selective medium, GFP visualization, and PCR analysis for the presence of the nptII gene, medium containing 200 mg/L kanamycin resulted in the highest transformation efficiency at 24%. This study sets the foundation for future genetic engineering and gene editing approaches for improvement of P. pruinosa.

scholarly journals Scaffolded Antigens in Yeast Cell Particle Vaccines Provide Protection against Systemic Polyoma Virus Infection

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
Donald J. Tipper ◽  
Eva Szomolanyi-Tsuda
Keyword(s):  
Yeast Cell ◽  
Large Scale ◽  
Fluorescent Protein ◽  
Subcutaneous Administration ◽  
Polyoma Virus ◽  
Apolipoprotein A1 ◽  
Yeast Cells ◽  
Scale Production ◽  
Protein Antigens ◽  
Virus Challenge

Background. U65, a self-aggregating peptide scaffold, traps fused protein antigens in yeast cells. Conversion to Yeast Cell Particle (YCP) vaccines by partial removal of surface mannoproteins exposesβ-glucan, mediating efficient uptake by antigen-presenting cells (APCs). YCP vaccines are inexpensive, capable of rapid large-scale production and have potential for both parenteral and oral use.Results. YCP processing by alkaline hydrolysis exposes up to 20% of the glucan but converts scaffolded antigen and internal yeast proteins into a common aggregate, preventing selective yeast protein removal. For U65-green fluorescent protein (GFP) or U65-Apolipoprotein A1 (ApoA1) subcutaneous vaccines, maximal IgG responses in mice required 10% glucan exposure. IgG responses to yeast proteins were 5-fold lower. Proteolytic mannoprotein removal produced YCPs with only 6% glucan exposure, insufficiently porous for selective removal of even native yeast proteins. Vaccine efficacy was reduced 10-fold. Current YCP formulations, therefore, are not suitable for human use but have considerable potential for use in feed animal vaccines. Significantly, a YCP vaccine expressing a GFP fusion to VP1, the murine polyoma virus major capsid protein, after either oral or subcutaneous administration, protected mice against an intraperitoneal polyoma virus challenge, reducing viral DNA levels in spleen and liver by >98%.

scholarly journals Expression, Purification, and Characterization of a Sucrose Nonfermenting 1-Related Protein Kinases 2 of Arabidopsis thaliana in E. coli-Based Cell-Free System

2016 ◽  
Vol 2016 ◽  
pp. 1-8
Author(s):  
Xu Zhang ◽  
Shuangxi Zhang ◽  
Jun Wang ◽  
Jing Zi ◽  
Jianhui Wang ◽  
...  
Keyword(s):  
Large Scale ◽  
Biochemical Properties ◽  
Plant Responses ◽  
Scale Production ◽  
Related Protein ◽  
Protein Preparation ◽  
Free System ◽  
Cell Free System ◽  
E Coli

The plant-specific sucrose nonfermenting 1-related protein kinase 2 (SnRK2) family is considered an important regulator of plant responses to abiotic stresses such as drought, cold, salinity, and nutrition deficiency. However, little information is available on how SnRK2s regulate sulfur deprivation responses in Arabidopsis. Large-scale production of SnRK2 kinases in vitro can help to elucidate the biochemical properties and physiological functions of this protein family. However, heterogenous expression of SnRK2s usually leads to inactive proteins. In this study, we expressed a recombinant Arabidopsis SnRK2.1 in a modified E. coli cell-free system, which combined two kinds of extracts allowing for a convenient and affordable protein preparation. The recombinant SnRK2.1 was produced in large-scale and the autophosphorylation activity of purified SnRK2.1 was characterized, allowing for further biochemical and substrate binding analysis in sulfur signaling. The application of this improved E. coli cell-free system provides us a promising and convenient platform to enhance expression of the target proteins economically.

scholarly journals Advanced Metabolic Engineering Approaches and Renewable Energy to Improve Environmental Benefits of Algal Biofuels: LCA of Large-scale Biobutanol Production with Cyanobacteria Synechocystis PCC6803

2021 ◽  
Author(s):  
Daniela Villacreses-Freire ◽  
Franziska Ketzer ◽  
Christine Rösch
Keyword(s):  
Genetic Engineering ◽  
Large Scale ◽  
Genetically Engineered ◽  
Environmental Benefits ◽  
Biofuel Production ◽  
Scale Production ◽  
Renewable Electricity ◽  
Synechocystis Pcc6803 ◽  
Cell Factories ◽  
Green Cell

AbstractWith modern genetic engineering tools, microorganisms can become resilient green cell factories to produce sustainable biofuels directly. Compared to non-engineered algae and cyanobacteria, the photon conversion efficiency can be significantly increased. Furthermore, simplified harvesting processes are feasible since the novel microorganisms are excreting the biofuels or their precursors continuously and directly into the cultivation media. Along with higher productivity and direct product harvesting, it is expected that environmental benefits can be achieved, especially for climate protection. A life cycle assessment (LCA) for biobutanol production with the genetically engineered cyanobacteria Synechocystis PCC6803 is performed to test this hypothesis. A prospective and upscaled approach was applied to assess the environmental impacts at large-scale production (20 ha plant) for better comparability with conventional butanol production. The LCA results show that the engineering of microorganisms can improve the environmental impact, mainly due to the higher productivity compared to non-engineered cyanobacteria. However, the nevertheless high electricity demand required for the cultivation and harvesting process overcompensates this benefit. According to the scenario calculations, a more favourable climate gas balance can be achieved if renewable electricity is used. Then, greenhouse gas emissions are reduced to 3.1 kg CO2 eq/kg biobutanol, corresponding to 20% more than the fossil reference: (2.45 kg CO2 eq./kg 1-butanol). The results indicate the importance of genetic engineering and the energy transition towards renewable electricity supply to take full advantage of the environmental potential of microorganisms as future green cell factories for sustainable biofuel production. Besides, the necessity of developing different scenarios for perspective and upscaled LCA for a fairer comparison with mature reference technologies is demonstrated.

AGRICULTURAL PRODUCTION OF CA DONG PEOPLE IN RESETTLEMENT AREA OF TRANH RIVER HYDROPOWER NUMBER 2: CURRENT SITUATION AND IMPACT FACTORS

2020 ◽  
Vol 9 (2) ◽  
Author(s):  
Bùi Thị Bích Lan
Keyword(s):  
Social Relations ◽  
Agricultural Production ◽  
Large Scale ◽  
Transition Process ◽  
Impact Factors ◽  
Small Scale ◽  
Scale Production ◽  
Technical Application ◽  
Production Models ◽  
Role Of The State

In Vietnam, the construction of hydropower projects has contributed significantly in the cause of industrialization and modernization of the country. The place where hydropower projects are built is mostly inhabited by ethnic minorities - communities that rely primarily on land, a very important source of livelihood security. In the context of the lack of common productive land in resettlement areas, the orientation for agricultural production is to promote indigenous knowledge combined with increasing scientific and technical application; shifting from small-scale production practices to large-scale commodity production. However, the research results of this article show that many obstacles in the transition process are being posed such as limitations on natural resources, traditional production thinking or the suitability and effectiveness of scientific - technical application models. When agricultural production does not ensure food security, a number of implications for people’s lives are increasingly evident, such as poverty, preserving cultural identity, social relations and resource protection. Since then, it has set the role of the State in researching and building appropriate agricultural production models to exploit local strengths and ensure sustainability.

I. S. A. Baud. Forms of Production and Women's Labour: Gender Aspects of Industrialisation in India and Mexico. New Delhi: Sage Publications. 1992. 321 pp.Hardbound. Price: Indian Rs 295.00.

1993 ◽  
Vol 32 (1) ◽  
pp. 129-131
Author(s):  
Naureen Talha
Keyword(s):  
Large Scale ◽  
Indian Society ◽  
New Delhi ◽  
Scale Production ◽  
Female Labour ◽  
Self Employment ◽  
Commodity Production ◽  
Large Scale Production ◽  
Mexican Society ◽  
Small Production

The literature on female labour in Third World countries has become quite extensive. India, being comparatively more advanced industrially, and in view of its size and population, presents a pictures of multiplicity of problems which face the female labour market. However, the author has also included Mexico in this analytical study. It is interesting to see the characteristics of developing industrialisation in two different societies: the Indian society, which is conservative, and the Mexican society, which is progressive. In the first chapter of the book, the author explains that he is not concerned with the process of industrialisation and female labour employed at different levels of work, but that he is interested in forms of production and women's employment in large-scale production, petty commodity production, marginal small production, and self-employment in the informal sector. It is only by analysis of these forms that the picture of females having a lower status is understood in its social and political setting.

Solid Lipid Nanoparticles: A Promising Drug Delivery Technology

2009 ◽  
Vol 2 (2) ◽  
pp. 509-516
Author(s):  
S. Pragati ◽  
S. Kuldeep ◽  
S. Ashok ◽  
M. Satheesh
Keyword(s):  
Drug Delivery ◽  
Solid Lipid Nanoparticles ◽  
Large Scale ◽  
Colloidal Particles ◽  
Scale Up ◽  
Lipid Nanoparticles ◽  
Scale Production ◽  
Research Activity ◽  
New Approach ◽  
Solid Lipid

One of the situations in the treatment of disease is the delivery of efficacious medication of appropriate concentration to the site of action in a controlled and continual manner. Nanoparticle represents an important particulate carrier system, developed accordingly. Nanoparticles are solid colloidal particles ranging in size from 1 to 1000 nm and composed of macromolecular material. Nanoparticles could be polymeric or lipidic (SLNs). Industry estimates suggest that approximately 40% of lipophilic drug candidates fail due to solubility and formulation stability issues, prompting significant research activity in advanced lipophile delivery technologies. Solid lipid nanoparticle technology represents a promising new approach to lipophile drug delivery. Solid lipid nanoparticles (SLNs) are important advancement in this area. The bioacceptable and biodegradable nature of SLNs makes them less toxic as compared to polymeric nanoparticles. Supplemented with small size which prolongs the circulation time in blood, feasible scale up for large scale production and absence of burst effect makes them interesting candidates for study. In this present review this new approach is discussed in terms of their preparation, advantages, characterization and special features.

Stepwise Development of Biomimetic Chimeric Peptides for Gene Delivery

2020 ◽  
Vol 27 (8) ◽  
pp. 698-710
Author(s):  
Roya Cheraghi ◽  
Mahboobeh Nazari ◽  
Mohsen Alipour ◽  
Saman Hosseinkhani
Keyword(s):  
Gene Delivery ◽  
Targeted Delivery ◽  
Viral Vectors ◽  
Large Scale ◽  
Transfection Efficiency ◽  
Cationic Lipids ◽  
Target Cells ◽  
Scale Production ◽  
Stepwise Development ◽  
Chimeric Peptides

Gene-based therapy largely relies on the vector type that allows a selective and efficient transfection into the target cells with maximum efficacy and minimal toxicity. Although, genes delivered utilizing modified viruses transfect efficiently and precisely, these vectors can cause severe immunological responses and are potentially carcinogenic. A promising method of overcoming this limitation is the use of non-viral vectors, including cationic lipids, polymers, dendrimers, and peptides, which offer potential routes for compacting DNA for targeted delivery. Although non-viral vectors exhibit reduced transfection efficiency compared to their viral counterpart, their superior biocompatibility, non-immunogenicity and potential for large-scale production make them increasingly attractive for modern therapy. There has been a great deal of interest in the development of biomimetic chimeric peptides. Biomimetic chimeric peptides contain different motifs for gene translocation into the nucleus of the desired cells. They have motifs for gene targeting into the desired cell, condense DNA into nanosize particles, translocate the gene into the nucleus and enhance the release of the particle into the cytoplasm. These carriers were developed in recent years. This review highlights the stepwise development of the biomimetic chimeric peptides currently being used in gene delivery.

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